Recombinant Mouse IL-17F

Recombinant Mouse IL-17F is used in research to study immune regulation, inflammation, host defense, and tissue remodeling, due to its role as a proinflammatory cytokine with distinct biological activities relevant to both health and disease models.

Key reasons to use recombinant mouse IL-17F in research applications include:

• Modeling Inflammatory Responses: IL-17F stimulates the production of proinflammatory cytokines such as IL-6, IL-8, and G-CSF, making it valuable for dissecting inflammatory signaling pathways and cytokine networks in vitro and in vivo.
• Studying Host Defense Mechanisms: IL-17F is implicated in mucosal immunity and defense against bacterial and viral infections by promoting neutrophil recruitment and activation, as well as enhancing antimicrobial peptide synthesis.
• Investigating Tissue Remodeling and Repair: IL-17F regulates cartilage matrix turnover by increasing matrix release and inhibiting new matrix synthesis, and it is involved in osteoblast maturation during bone repair.
• Exploring Angiogenesis and Fibrosis: IL-17F can inhibit angiogenesis and induce endothelial cells to produce factors such as IL-2, TGF-β, and MCP-1, which are relevant for studies on vascular biology and fibrotic processes.
• Elucidating Disease Pathogenesis: IL-17F plays roles in autoimmune diseases, allergic responses, asthma, and colitis, making it a target for mechanistic studies and therapeutic intervention research.
• Comparative Studies with IL-17A: IL-17F shares some functions with IL-17A but also has unique roles, allowing researchers to dissect the specific contributions of each cytokine in immune regulation and pathology.

Typical applications include:

• Cell-based bioassays to measure cytokine induction or cell migration.
• In vivo models of inflammation, infection, allergy, or tissue injury.
• Mechanistic studies of cytokine signaling and gene regulation.
• Screening or validation of anti-inflammatory or immunomodulatory compounds.

Using recombinant mouse IL-17F ensures reproducibility and specificity in experimental systems, as it provides a defined, biologically active cytokine for controlled stimulation or inhibition studies.

Yes, recombinant mouse IL-17F can be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity, properly quantified, and compatible with your assay system. This is a common practice in cytokine ELISAs, where recombinant proteins serve as standards to generate a calibration curve for quantifying unknown samples.

Key considerations and best practices:

• Purity and Quantification: Ensure the recombinant IL-17F is highly pure and accurately quantified. The standard should be reconstituted and diluted according to the manufacturer's or protocol's instructions to achieve precise concentrations for the standard curve.
• Formulation: Recombinant IL-17F is often available in carrier-free or BSA-containing formulations. For ELISA standards, BSA-containing formulations are generally preferred for stability, unless BSA interferes with your assay.
• Validation: The recombinant standard should be validated for use in your specific ELISA system. Most commercial ELISA kits use recombinant IL-17F as the standard and provide detailed protocols for its preparation and use.
• Standard Curve Preparation: Prepare a serial dilution of the recombinant IL-17F to generate a standard curve covering the expected concentration range in your samples (e.g., 3.13–200 pg/mL).
• Matrix Effects: If quantifying IL-17F in complex matrices (e.g., serum, plasma), confirm that the recombinant standard behaves similarly to native IL-17F in your assay. Recovery and linearity experiments are recommended to ensure accuracy.
• Batch Consistency: Recombinant proteins produced using standardized methods offer high batch-to-batch consistency, which is critical for reproducible quantification.

Limitations:

• The recombinant standard should match the native protein in terms of immunoreactivity. Most commercial recombinant IL-17F proteins are validated for this purpose, but if using a custom preparation, confirm its suitability.
• Some recombinant proteins are not tested for bioactivity in functional assays, but this does not affect their use as ELISA standards.

Summary Table: Use of Recombinant Mouse IL-17F as ELISA Standard

In summary: Recombinant mouse IL-17F is widely used and appropriate as a standard for ELISA quantification, provided it is validated for your assay and handled according to best practices.

Recombinant Mouse IL-17F has been validated for several key applications in published research, primarily in bioassays and in vivo studies investigating immune function, inflammation, and disease models.

Validated Applications in Published Research:

• Bioassay (in vitro functional studies):

  • Used to stimulate cells (e.g., T cells, monocytes, endothelial cells, fibroblasts) to assess cytokine production (such as IL-6, IL-8, G-CSF), chemokine induction, and regulation of matrix turnover.
  • Employed in studies of osteoblast maturation, colitis protection via Treg induction, and host defense against bacterial infection.
  • Applied to characterize selective neutrophil attraction in models of psoriasis and airway inflammation.

• In Vivo Studies (animal models):

  • Used to investigate the role of IL-17F in allergic airway sensitization, neutrophilia, airway hyperresponsiveness, and pulmonary inflammation.
  • Validated in models of fracture repair, hepatitis, and immune responses to infection.
  • Applied in studies of autoimmune and chronic inflammatory diseases such as rheumatoid arthritis and multiple sclerosis.

• Cell Culture:

  • Utilized as a supplement in cell culture systems to study cytokine signaling, immune cell differentiation, and chemokine production.

• ELISA Standard:

  • Used as a standard for quantifying IL-17F in immunoassays, though this is a technical validation rather than a direct research application.

Representative Published Studies:

• Colitis and Treg induction: Recombinant Mouse IL-17F was used to demonstrate that suppression of IL-17F protects against colitis by modifying intestinal microbiota and promoting regulatory T cell induction.
• Osteoblast maturation: Validated for studying T-lymphocyte-mediated osteoblast maturation during fracture repair.
• Airway inflammation: Used in vivo to model Th17-dependent neutrophilia and airway hyperresponsiveness following allergic sensitization.
• Psoriasis: Functional characterization as a selective neutrophil attractant in skin inflammation models.
• Host defense: Differential roles of IL-17A and IL-17F in defense against mucoepithelial bacterial infection.

Summary Table of Validated Applications

Additional Notes:

• IL-17F is often studied alongside IL-17A due to overlapping and distinct biological functions.
• It is used to dissect mechanisms of inflammation, immune cell recruitment, and tissue remodeling in both health and disease contexts.
• Recombinant Mouse IL-17F is not typically validated for diagnostic or therapeutic use in humans; its applications are restricted to preclinical and basic research settings.

If you require protocols or specific experimental details for any of these applications, please specify the context or research focus.

To reconstitute and prepare Recombinant Mouse IL-17F protein for cell culture experiments, follow these best-practice steps based on current protocols:

1. Centrifuge the vial:
Before opening, briefly centrifuge the vial (e.g., 3000 rpm for 5 minutes) to ensure all lyophilized protein is at the bottom.

2. Reconstitution:

• Add sterile water or the recommended buffer to achieve a concentration of at least 100 μg/mL.
• If the product datasheet specifies, use sterile 4 mM HCl (with or without 0.1% BSA or HSA as a carrier protein) for reconstitution.
• Gently pipette to dissolve. Do not vortex or shake vigorously, as this may denature the protein and reduce biological activity.
• Allow the solution to sit at room temperature for 15–20 minutes to ensure complete dissolution.

3. Dilution for cell culture:

• After reconstitution, further dilute the protein to your working concentration using cell culture medium or a buffer containing a stabilizer such as 0.1% BSA, 10% FBS, 5% HSA, or 5% trehalose.
• Prepare dilutions fresh or aliquot and freeze for longer-term storage.

4. Storage:

• Lyophilized protein: Store at –20°C or lower for long-term stability.
• Reconstituted protein: Store at 2–8°C for up to 1 week. For longer storage, aliquot and freeze at –20°C or –80°C. Avoid repeated freeze/thaw cycles.

5. Additional notes:

• The protein is typically supplied carrier-free or with minimal additives. If your application is sensitive to carrier proteins, confirm the formulation before use.
• For functional assays, confirm the activity (e.g., induction of IL-6 in responsive cell lines) as described in the product documentation.

Summary protocol example:

1. Centrifuge vial at 3000 rpm for 5 min.2. Add sterile H₂O or 4 mM HCl (with 0.1% BSA if required) to reach 100 μg/mL.3. Gently pipette to dissolve; do not vortex.4. Incubate at room temperature for 15–20 min.5. Dilute to working concentration in cell culture medium with stabilizer.6. Store aliquots at –20°C or –80°C for long-term use.

Key precautions:

• Do not vortex the protein solution.
• Use sterile technique throughout.
• Avoid repeated freeze/thaw cycles to preserve activity.

These steps are consistent with protocols from multiple suppliers and are suitable for most cell culture applications involving recombinant mouse IL-17F. Always consult the specific product datasheet for any unique requirements.